Fermentation, Volume 5, Issue 3 (September 2019) – 32 articles

Fusarium head blight (FHB) is one of the most significant causes of economic loss in cereal crops, resulting in a loss of $50–300 million for Canadian agriculture. The infected grain (containing Fusarium-damaged kernels (FDKs)) is often both lower in quality and kernel weight, and it may be unsuitable for human and animal consumption due to mycotoxin presence. However, it still contains a considerable amount of nutrients. A method to recover the nutrients without the mycotoxins should be beneficial for the agricultural economy. In this study, our objective was to examine recovery methods of the nutrients in relation to mycotoxin accumulation in the insect. The FDKs were fermented with Aspergillus oryzae and/or Lactobacillus plantarum (solid-state fermentation (SSF)). The SSF kernels were then provided to 50 young, black soldier fly larvae (BSFL) for 12 days. Weight gain, chemical composition, and mycotoxin bioaccumulation of BSFL and spent feed were evaluated. After 12 days of insect culture, the BSFL grew 5–6 times their initial weight. While the overall weights did not significantly vary, the proteins and lipids accumulated more in SSF FDK-fed insects. During the active growth period, the larval biomass contained deoxynivalenol (DON), a mycotoxin, at detectable levels; however, by day 12, when the larvae were in the pre-pupal stage, the amount of DON in the insect biomass was nearly negligible, i.e., BSFL did not accumulate DON. Thus, we conclude that the combination of BSFL and SSF can be employed to recover DON-free nutrients from FHB-infected grain to recover value from unmarketable grain. Full article

Anhydrobiosis is a unique state of live organisms in which their metabolism is temporary reversibly suspended as the result of strong dehydration of their cells. This state is widely used currently during large-capacity production of active dry baker’s yeast. Other strains of the yeast Saccharomyces cerevisiae, as well as other yeast species that could potentially find use in modern biotechnology, are not resistant to dehydration–rehydration treatments. To improve their resistance, the main factors that influence cell survival during such treatment need to be revealed. This study showed the importance of mitochondria for yeast cell survival during transfer into anhydrobiosis, a factor that was strongly underestimated until this study. It was revealed that the external introduction inside yeast cells of 50 μM of lithocholic acid (LCA), an agent that induces changes in glycerophospholipids in mitochondrial membranes, in combination with 1% DMSO, may improve the survival rate of dehydrated cells. The influence of LCA upon yeast cell resistance to dehydration–rehydration was not linked with changes in the state of the cells’ plasma membrane. Full article

About one-third of the food produced annually worldwide ends up as waste. A minor part of this waste is used for biofuel and compost production, but most is landfilled, causing environmental damage. Mass production of edible insects for human food and livestock feed seems a sustainable solution to meet demand for animal-based protein, which is expected to increase due to rapid global population growth. The aim of this review was to compile up-to-date information on mass rearing of edible insects for food and feed based on food wastes. The use and the potential role of the fermentation process in edible insect mass production and the potential impact of this rearing process in achieving an environmentally friendly and sustainable food industry was also assessed. Food waste comprises a huge nutrient stock that could be valorized to feed nutritionally flexible edible insects. Artificial diets based on food by-products for black soldier fly, house fly, mealworm, and house cricket mass production have already been tested with promising results. The use of fermentation and fermentation by-products can contribute to this process and future research is proposed towards this direction. Part of the sustainability of the food sector could be based on the valorization of food waste for edible insect mass production. Further research on functional properties of reared edible insects, standardization of edible insects rearing techniques, safety control aspects, and life cycle assessments is needed for an insect-based food industry. Full article

This research aimed to evaluate the impact of different options for winemaking on the color composition of Uruguayan Tannat red wines. The techniques evaluated were the substitution of ripe grape juice with immature grape juice and the heating of the crushed grapes before fermentation, called must replacement and hot pre-fermentative maceration, respectively. These procedures were proposed to reduce the alcohol content and increase the phenolic composition of the wine, according to the expected effects of climate change and current trends in consumer preferences. The investigation was made over three consecutive years (2016, 2017, and 2018). Both winemaking techniques allow the enhancement of the chromatic characteristics of wines via the modification of the phenolic composition. Additionally, such techniques allow the overcoming of the well-known limitations in the extractability of anthocyanins presented by the Tannat cultivar. Hot pre-fermentative maceration increases the proportion of the most oxidizable molecules delphinidin-3-O-glucoside, cyanidin-3-O-glucoside, and petunidin-3-O-glucoside, suggesting heat inactivation of polyphenoloxidases enzymes. Must replacement and hot pre-fermentative maceration are technological alternatives that could significantly improve the intensity and chromatic characteristics of red wines. Full article

In recent years, studies have reported the positive influence of non-Saccharomyces yeast on wine quality. Many grape varieties under mixed or sequential inoculation show an overall positive effect on aroma enhancement. A potential impact by non-Saccharomyces yeast on volatile and non-volatile compounds should benefit the flavor of Riesling wines. Following this trend, four separate sequential fermentations (using the non-Saccharomyces yeasts Torulaspora delbrueckii, Metschnikowia pulcherrima, Pichia kluyveri, and Lachancea thermotolerans with Saccharomyces cerevisiae) were carried out on Riesling must and compared to a pure culture of S. cerevisiae. Sequential fermentations influenced the final wine aroma. Significant differences were found in esters, acetates, higher alcohols, fatty acids, and low volatile sulfur compounds between the different trials. Other parameters, including the production of non-volatile compounds, showed significant differences. This fermentation process not only allows the modulation of wine aroma but also chemical parameters such as glycerol, ethanol, alcohol, acidity, or fermentation by-products. These potential benefits of wine diversity should be beneficial to the wine industry. Full article

In winemaking processes, there is a current tendency to develop spontaneous fermentations taking advantage of the metabolic diversity of derived from the great microbial diversity present in grape musts. This enological practice enhances wine complexity, but undesirable consequences or deviations could appear on wine quality. Soil is a reservoir of important microorganisms for different beneficial processes, especially for plant nutrition, but it is also the origin of many of the phytopathogenic microorganisms that affect vines. In this study, a meta-taxonomic analysis of the microbial communities inhabiting vineyard soils was realized. A significant impact of the soil type and climate aspects (seasonal patterns) was observed in terms of alpha and beta bacterial diversity, but fungal populations appeared as more stable communities in vineyard soils, especially in terms of alpha diversity. Focusing on the presence and abundance of wine-related microorganisms present in the studied soils, some seasonal and soil-dependent patterns were observed. The Lactobacillaceae family, containing species responsible for the malolactic fermentation, was only present in non-calcareous soils samples and during the summer season. The study of wine-related fungi indicated that the Debaryomycetaceae family dominates the winter yeast population, whereas the Saccharomycetaceae family, containing the most important fermentative yeast species for winemaking, was detected as dominant in summer. Full article

The goal of this study was to examine cost impacts using techno-economic analysis (TEA) and environmental impacts using life-cycle assessment (LCA) for the production of red wine. Three production scales, denoted as “small” (5000 gal per year), “medium” (50,000 gal per year), and “large” (500,000 gal per year) were chosen for analysis. For example, the consumption of water, energy, greenhouse gas emissions, and solid waste generation were considered in order to estimate environmental impacts. A spreadsheet-based economic model was also developed. The results of the LCA and TEA were compared amongst all production scales. The results of the LCA showed that both bottle manufacturing and various wine-making processes contributed the greatest environmental impacts. For TEA, the relationships between costs and profits increased as production scale increased; exponential trend lines could describe the data, but linear models were better. This information can be useful when considering what size of winery might be appropriate to invest in, or what operational categories may be most impactful in terms of costs and environmental burdens and, thus, may be targets for efficiency improvements. Full article

The degradation dynamics of lignin and cellulose were analyzed by means of a solid state biodegradation experiment, using residues from the essential oil extraction of the Palo Santo tree (Bursera graveolens). As such, two native Xylaria spp. and an exotic mushroom Trametes versicolor were incubated on the spent substrate (Residues of B. Graveolens, BGR’s). The relatively high lignin and cellulose contents of the BGRs (9.1% and 19%, respectively) indicated the potential of this resource for the production of methane (biogas) and ethanol. However, the degradation of the lignin and cellulose content could be traced back to the relatively high activity of the enzymes laccase, cellulase, and xylanase, produced by the fungi. The results showed that laccase (30.0 U/L and 26.6 U/L), cellulase (27.3 U/L and 35.8 U/L) and xylanase (189.7U/L and 128.3 U/L) activities of Xylaria feejeensis and Xylaria cf. microceras were generally higher than T. versicolor (9.0 U/L, 29.5 U/L, 99.5 U/L respectively). Furthermore, the total carbon (TC: 47.3%), total nitrogen (TN: 1.5%), total phosphorus (TP: 0.2%) and total potassium (TK: 1.2%) dynamics were analyzed during the experiment and their importance for the degradation process highlighted. The results of this work might serve as guidance for future studies in dry forest areas, while furthering the understanding of the potential use of native fungi as ecologic lignocellulosic decomposers and for industrial proposes. Full article

Syngas (mixture of CO, H₂ and CO₂) fermentation suffers from mass transfer limitation due to low solubility of CO and H₂ in the liquid medium. Therefore, it is critical to characterize the mass transfer in syngas fermentation reactors to guide in delivery of syngas to the microorganisms. The objective of this study is to measure and predict the overall volumetric mass transfer coefficient, k_La for O₂ at various operating conditions in a 7-L sparged and non-sparged continuous stirred-tank reactor (CSTR). Measurements indicated that the k_La for O₂ increased with an increase in air flow rate and agitation speed. However, k_La for O₂ decreased with the increase in the headspace pressure. The highest k_La for O₂ with air sparged in the CSTR was 116 h⁻¹ at 600 sccm, 900 rpm, 101 kPa, and 3 L working volume. Backmixing of the headspace N₂ in the sparged CSTR reduced the observed k_La. The mass transfer model predicted the k_La for O₂ within 10% of the experimental values. The model was extended to predict the k_La for syngas components CO, CO₂ and H₂, which will guide in selecting operating conditions that minimize power input to the bioreactor and maximize the syngas conversion efficiency. Full article

Three yeasts (Saccharomyces cerevisiae var. boulardii, a commercial probiotic yeast; S. cerevisiae W13, a wild yeast able to remove ochratoxin A; and S. cerevisiae 17, a wild yeast with promising probiotic traits) were screened for their ability to adhere on apple pieces as a function of different contact times (15–30 min). Then, apple pieces were stored at 4 °C for 15 days, and the viable count of yeasts was periodically assessed. Yeasts were able to adhere on apple pieces after 15 min (7 log cfu/g) and retained their viability throughout the refrigerated storage. In a second step, apple pieces with S. cerevisiae W13 were used to produce cider on a small scale. The variables under investigation were (a) the recycling of pieces up to 10 times and (b) the preliminary storage of pieces at 4 °C before use. Pieces used immediately after yeast immobilization could be successfully used again 10 times and gained a fermentation performance (in terms of yeast amount in cider and ethanol after 24 h) similar to that achieved by free cells. In addition, the preliminary storage of pieces at 4 °C did not affect their performances as reusable starter carriers. Full article

(1) Background: It has been shown that d-galacturonic acid is converted to l-galactonic acid by the basidiomycotic yeast, Cryptococcus diffluens. However, two pathways are hypothesized for the l-galactonic acid conversion process in C. diffluens. One is similar to the conversion process of the filamentous fungi in d-galacturonic acid metabolism and another is the conversion process to l-ascorbic acid, reported in the related yeast, C. laurentii. It is necessary to determine which, if either, process occurs in C. diffluens in order to produce novel value-added products from d-galacturonic acid using yeast strains. (2) Methods: The diethylaminoethy (DEAE)-fractionated enzyme was prepared from the cell-free extract of C. diffluens by the DEAE column chromatography. The l-galactonic acid conversion activity was assayed using DEAE-fractionated enzyme and the converted product was detected and fractionated by high-performance anion-exchange chromatography. Then, the molecular structure was identified by nuclear magnetic resonance analysis. (3) Results: The product showed similar chemical properties to 2-keto-3-deoxy-l-galactonic acid (l-threo-3-deoxy-hexulosonic acid). (4) Conclusions: It is suggested that l-galactonic acid is converted to 2-keto-3-deoxy-l-galactonic acid by dehydratase in C. diffluens. The l-galactonic acid conversion process of C. diffluens is a prioritized pathway, similar to the pathway of ascomycetes. Full article

Cashew apples are by-products in the production of cashew nuts, which are mostly left to rot in the fields. Cashew apple juice (CAJ), a highly nutritious beverage, can be produced from them. It is rich in sugars and ascorbic acid, but its high polyphenol content makes it bitter and astringent, and therefore difficult to commercialize. The kingdom of fungi contains more than 2000 yeast species, of which only a few species have been studied in relation to their potential to produce aroma compounds. The aim of this research was to develop a new low-alcoholic fermented beverage to valorize cashew apples. For this purpose, a screening was carried out employing non-conventional yeast species and some species of the genus Saccharomyces for comparison, followed by a more detailed study with four selected strains cultured at different conditions. The production of volatile aroma compounds as a function of the presence of oxygen, temperature, and yeast species was investigated. The results showed that the more diverse aroma profiles appeared at 25 °C under anaerobic cultivation conditions, where Saccharomyces cerevisiae WUR 102 and Hanseniaspora guilliermondii CBS 2567 excelled in the synthesis of certain aroma compounds, such as β-phenylethanol and its acetate ester (rose aroma). Further studies are needed to test consumer acceptance of these new products. Full article

Vitis vinifera cv. Narince is a Turkish native white grape variety. In this study, volatile and sensory properties of Narince wines that are produced with autochthonous Saccharomyces cerevisiae (S. cerevisiae) strains and commercial strain were compared. Autochthonous yeast strains 1044 (MG017575), 1088 (MG017577), and 1281 (MG017581) were previously isolated from spontaneous fermentations of Narince grapes. Volatile compounds formed in wines were extracted using a liquid–liquid extraction method and determined by GC-MS-FID. All yeast strains fermented Narince grape juice to dryness. The differences between the volatile profiles of the yeast strains were determined. Wines fermented with autochthonous strains 1281 and 1044 produced a higher amount of acetates and ethyl esters. While the highest concentrations of ethyl hexanoate and hexyl acetate were found in wine fermented with 1044, the highest concentrations of ethyl octanoate, ethyl decanoate, isoamyl acetate, and 2-phenylethyl acetate were found in wine fermented with strain 1281. Also, the highest contents of 2-phenyl ethanol and linalool were found in wine fermented with strain 1281. According to sensory analysis, the wine fermented with 1281 achieved the best scores in floral and fruity attributes, as well as balance and global impression. The data obtained in the present study showed that autochthonous yeast strains affect the final physicochemical composition and sensory profile of Narince wines. Full article

Sulfur dioxide (SO₂) is an antioxidant and antimicrobial agent used in winemaking. Its effects on spoilage microorganisms has been studied extensively, but its effects on commercial Saccharomyces cerevisiae strains, the dominant yeast in winemaking, require further investigation. To our knowledge, no previous studies have investigated both the potential SO₂ resistance mechanisms of commercial yeasts as well as their production of aroma-active volatile compounds in response to SO₂. To study this, fermentations of two commercial yeast strains were conducted in the presence (50 mg/L) and absence (0 mg/L) of SO₂. Strain QA23 was more sensitive to SO₂ than Strain BRL97, resulting in delayed cell growth and slower fermentation. BRL97 exhibited a more rapid decrease in free SO₂, a higher initial production of hydrogen sulfide, and a higher production of acetaldehyde, suggesting that each strain may utilize different mechanisms of sulfite resistance. SO₂ addition did not affect the production of aroma-active volatile compounds in QA23, but significantly altered the volatile profiles of the wines fermented by BRL97. Full article

Removal of the mucilage layer of coffee fruits by a fermentation process has became an interesting strategy to improve coffee quality, which is able to assist the formation of flavored molecules. In this study, four sets of inoculation protocols were evaluated using ripe and immature coffee fruits, respectively, including (i) pure culture fermentation with Pichia fermentans, (ii) pure culture fermentation with Pediococcus acidilactici, (ii) combined fermentation with P. fermentans and P. acidilactici, and (iv) spontaneous, non-inoculated control. The initial pulp sugar concentration of ripe coffee fruits (0.57 and 1.13 g/L glucose and fructose content, respectively) was significantly higher than immature coffee pulp (0.13 and 0.26 g/L glucose and fructose content, respectively). Combined inoculation with P. fermentans and P. acidilactici of ripe coffee beans increased pulp sugar consumption and production of metabolites (lactic acid, ethanol, and ethyl acetate), evidencing a positive synergic interaction between these two microbial groups. On the other hand, when immature coffee fruits were used, only pure culture inoculation with P. fermentans was able to improve metabolite formation during fermentation, while combined treatment showed no significant effect. Altogether, 30 volatile compounds were identified and semi-quantified with HS- solid phase microextraction (SPME)-gas chromatography coupled to mass spectrophotometry (GC/MS) in fermented coffee beans. In comparison with pure cultures and spontaneous process, combined treatment prominently enhanced the aroma complexity of ripe coffee beans, with a sharp increase in benzeneacetaldehyde, 2-heptanol, and benzylalcohol. Consistent with the monitoring of the fermentation process, only P. fermentans treatment was able to impact the volatile composition of immature coffee beans. The major impacted compounds were 2-hexanol, nonanal, and D-limonene. In summary, this study demonstrated the great potential of the combined use of yeast and lactic acid bacteria to improve fermentation efficiency and to positively influence the chemical composition of coffee beans. Further studies are still required to investigate the mechanisms of synergism between these two microbial groups during the fermentation process and influence the sensory properties of coffee products. Full article

The production of volatile compounds has become one of the major technological features for yeast selection. In fact, although the aromatic profile of the wine is the sum of varietal-, pre-, post-, and fermentative-aroma compound, yeasts affect the quality of the grape from maturation throughout fermentation, metabolizing sugars and other components into alcohols, esters, organic acids, and aldehydes. Among the new technological features, the production of mannoproteins has gained interest. From this perspective, the main aim of this work was to characterize 9 strains of Saccharomyces cerevisiae and 1 of Saccharomyces bayanus for their volatile profiles and the release of mannoproteins. The strains were inoculated in Trebbiano musts and incubated at 15 °C; at the end of fermentation the wines were evaluated by GC/MS/SPME for their volatile profiles and mannoprotein content by enzymatic assay. The strains were inoculated at level ranging between 4.9 and 6.3 log CFU/mL but only the strains L318 and 12233X6167 were able to reach values of 7.5 log CFU/mL. The aromatic profiles resulted in a strain-specific fingerprinting. According to the principal component analysis, the wines produced by the strains L288, L234, and L318 were characterized by the presence of propanoic acid, butanol, octanoic acid, and 3 methyl pentanol while the wine obtained by the strain 12233x35G2 was characterized by the presence of propanoic acid, butanol, octanoic acid and 3 methyl pentanol while the strain 12233x35G2 was characterized by the presence of decanoic acid ethyl ester, heptanoic acid ethyl ester, and acetic acid 2 phenetyl ester. Regarding mannoproteins, the highest concentration was achieved by strain12233x6167 (104 mg/L). The data allowed to select the strains endowed with the best fermentation performances in terms of aroma and mannoproteins release. Full article

It is well known that high ethanol levels in wines adversely affect the perception of new wine consumers. Moreover, numerous issues, such as civil restrictions, health risk and trade barriers, are associated with high ethanol concentrations. Several strategies have been proposed to produce wines with lower alcoholic content, one simple and inexpensive approach being the use of new wine native yeasts with less efficiency in sugar to ethanol conversion. Nevertheless, it is also necessary that these yeasts do not impair the quality of wine. In this work, we tested the effect of sequential culture between Hanseniaspora uvarum BHu9 and Saccharomyces cerevisiae BSc114 on ethanol production. Then, the wines produced were analyzed by GC-MS and tested by a sensorial panel. Co-culture had a positive impact on ethanol reduction and sensory profile when compared to the S. cerevisiae monoculture. Wines with lower alcohol content were related to fruity aroma; moreover, color intensity was associated. The wines obtained with S. cerevisiae BSc114 in pure conditions were described by parameters linked with high ethanol levels, such as hotness and astringency. Moreover, floral profile was related to this treatment. Based on these findings, this work provides a contribution to answer the current consumers’ preferences and addresses the main challenges faced by the enological industry. Full article

Hanseniaspora uvarum is one of the predominant non-Saccharomyces yeast species found on grapes and in juice, but its effect on lactic acid bacteria (LAB) growth and wine flavor has not been extensively studied. Therefore, the interaction between H. uvarum, two Saccharomyces cerevisiae yeast strains, two LAB species (Lactobacillus plantarum and Oenococcus oeni) in combination with two malolactic fermentation (MLF) strategies was investigated in Shiraz wine production trials. The evolution of the different microorganisms was monitored, non-volatile and volatile compounds were measured, and the wines were subjected to sensory evaluation. Wines produced with H. uvarum in combination with S. cerevisiae completed MLF in a shorter period than wines produced with only S. cerevisiae. Sequential MLF wines scored higher for fresh vegetative and spicy aroma than wines where MLF was induced as a simultaneous inoculation. Wines produced with H. uvarum had more body than wines produced with only S. cerevisiae. The induction of MLF using L. plantarum also resulted in wines with higher scores for body. H. uvarum can be used to reduce the duration of MLF, enhance fresh vegetative aroma and improve the body of a wine. Full article

Metschnikowia pulcherrima (Mp) is a ubiquitous yeast that frequently appears in spontaneous fermentations. The current interest in Mp is supported by the expression of many extracellular activities, some of which enhance the release of varietal aromatic compounds. The low fermentative power of Mp makes necessary the sequential or mixed use with Saccharomyces cerevisiae (Sc) to completely ferment grape musts. Mp has a respiratory metabolism that can help to lower ethanol content when used under aerobic conditions. Also, Mp shows good compatibility with Sc in producing a low-to-moderate global volatile acidity and, with suitable strains, a reduced level of H₂S. The excretion of pulcherrimin gives Mp some competitive advantages over other non-Saccharomyces yeasts as well as providing some antifungal properties. Full article

In this study, two vineyards of different age were chosen. During three years, a sampling campaign was performed for isolating vineyard-associated Saccharomyces cerevisiae (S. cerevisiae) strains. Bark portions and, when present, grape bunches were regularly collected from the same vine plants during the overall sampling period. Each bark portion was added to a synthetic must, while each grape bunch was manually crushed, and fermentations were run to isolate S. cerevisiae strains. All collected yeasts were identified at different species and strain levels to evaluate the genetic variability of S. cerevisiae strains in the two vineyards and strains dynamics. Moreover, bark-associated strains were compared with those isolated from spontaneous fermentations of grapes collected during the two harvests. Regarding the youngest vineyard, no S. cerevisiae was identified on bark and grape surface, highlighting the importance of vine age on yeast colonization. Results reported the isolation of S. cerevisiae from vine bark of the old vineyard at all sampling times, regardless of the presence of the grape bunch. Therefore, this environment can be considered an alternative ecological niche that permanently hosts S. cerevisiae. Bark-associated strains were not found on grape bunches and during pilot-scale vinifications, indicating no significative strain transfer from vine bark to the grape must. Commercial starters were identified as well both in vineyards and during vinifications. Full article

This study is focused on using multi-criteria decision making (MCDM) for selecting specific growth rate models of batch cultivation by the Saccharomyces cerevisiae. Ten specific growth rate models—Monod, Mink, Tessier, Moser, Aiba, Andrews, Haldane, Luong, Edward, and Han-Levenspiel—were investigated in order to explain the cell growth kinetics by the dependence on glucose. By using the preference ranking organization method (PROMETHEE) II, it was found that the Andrews model was the highest of rank and was the most appropriate one for modelling. Full article

Two major spoilage yeasts in the wine industry, Brettanomyces bruxellensis and Zygosaccharomyces rouxii, produce off-flavors and gas, causing considerable economic losses. Traditionally, SO₂ has been used in winemaking to prevent spoilage, but strict regulations are in place regarding its use due to its toxic and allergenic effects. To reduce its usage researchers have been searching for alternative techniques. One alternative is biocontrol, which can be used either independently or in a complementary way to chemical control (SO₂). The present study analyzed 122 native non-Saccharomyces yeasts for their biocontrol activity and their ability to be employed under fermentation conditions, as well as certain enological traits. After the native non-Saccharomyces yeasts were assayed for their biocontrol activity, 10 biocontroller yeasts were selected and assayed for their ability to prevail in the fermentation medium, as well as with respect to their corresponding positive/negative contribution to the wine. Two yeasts that satisfy these characteristics were Wickerhamomyces anomalus BWa156 and Metschnikowia pulcherrima BMp29, which were selected for further research in application to mixed fermentations. Full article

The study and use of non-Saccharomyces yeasts to wine improvement and diversification has gained considerable relevance in recent years. The present work reports a pilot-scale winery assay of mixed fermentation with a commercial strain of Metschnikowia pulcherrima, tested in five white and nine red grape varieties. Two modalities were assayed, one with the addition of M. pulcherrima at time zero and addition of Saccharomyces cerevisiae after 24 h, and a control using only S. cerevisiae at time zero. Fermentation was monitored by daily measurement of density and temperature. Wine physicochemical analysis was performed after winemaking and repeated after four years of aging. Variance and multivariate analysis were used to examine these data. Triangle and ranking tests were performed on the wines obtained, using an experienced sensory panel. Alcoholic fermentation proceeded smoothly until there was complete consumption of the sugars. M. pulcherrima in mixed fermentation, although mainly recommended for white wine, was also tested for red wines. These wines generally presented higher glycerol, reducing sugars and total dry matter, and lower alcohol content, in line with the current market trend. Significant sensory differences among modalities were only obtained for three varieties. Results emphasized that grape variety is a relevant factor in studies with non-Saccharomyces yeasts. Full article

Nitrogen is a limiting factor for the development of wine alcoholic fermentation. The addition of nutrients and different nitrogen sources is a usual practice for many winemakers. Currently, there is a market trend toward wine that is additive-free and there are also restrictions on the amount of ammonium fermentation agents that can be added to the wine. In this work, the changes produced on the alcoholic fermentation by the addition of different nitrogen sources were evaluated by the use of ATR-FTIR. The results showed the feasibility of this technique to observe differences in the growth yeast capacity depending on the type of the nutrients added. A high influence on the development of the alcoholic fermentation was observed, especially at its exponential and the stationary phases. Moreover, the changes observed in the recorded spectra were related to the proteins and lipid esters composition of the yeast cell wall. This technique should be a useful tool to evaluate nitrogen deficiencies during winemaking although further studies should be done in order to evaluate more influential factors. Full article

Under anaerobic/hypoxic conditions, Saccharomyces cerevisiae relies on external lipid supplements to modulate membrane lipid fraction in response to different stresses. Here, transcriptomic responses of two S. cerevisiae wine strains were evaluated during hypoxic fermentation of a synthetic must with/without ergosterol and oleic acid supplementation. In the absence of lipids, the two strains, namely EC1118 and M25, showed different behaviour, with M25 significantly decreasing its fermentation rate from the 72 h after inoculum. At this time point, the whole genome transcriptomic analysis revealed common and strain-specific responses to the lack of lipid supplementation. Common responses included the upregulation of the genes involved in ergosterol biosynthesis, as well as the seripauperin and the heat shock protein multigene families. In addition, the upregulation of the aerobic isoforms of genes involved in mitochondrial electron transport is compatible with the previously observed accumulation of reactive oxygen species in the two strains during growth in absence of lipids. Considering the strain-specific responses, M25 downregulated the transcription of genes involved in glucose transport, methionine biosynthesis and of those encoding mannoproteins required for adaptation to low temperatures and hypoxia. The identification of these pathways, which are presumably involved in yeast resistance to stresses, will assist industrial strain selection. Full article

In this contribution, the effect of the presence of a presumed inert gas like ${\mathrm{N}}_2$ in the feed gas on the biological methanation of hydrogen and carbon dioxide with Methanothermobacter marburgensis was investigated. ${\mathrm{N}}_2$ can be found as a component besides ${\mathrm{CO}}_2$ in possible feed gases like mine gas, weak gas, or steel mill gas. To determine whether there is an effect on the biological methanation of ${\mathrm{CO}}_2$ and ${\mathrm{H}}_2$ from renewable sources or not, the process was investigated using feed gases containing ${\mathrm{CO}}_2$ , ${\mathrm{H}}_2$ , and ${\mathrm{N}}_2$ in different ratios, depending on the ${\mathrm{CO}}_2$ content. A possible effect can be a lowered conversion rate of ${\mathrm{CO}}_2$ and ${\mathrm{H}}_2$ to ${\mathrm{CH}}_4$ . Feed gases containing up to 47% ${\mathrm{N}}_2$ were investigated. The conversion of hydrogen and carbon dioxide was possible with a conversion rate of up to 91% but was limited by the amount of ${\mathrm{H}}_2$ when feeding a stoichiometric ratio of 4:1 and not by adding ${\mathrm{N}}_2$ to the feed gas. Full article

The characteristics of fermented milk are affected by the type of milk used and the microorganisms involved in the fermentation process. Goat milk has been widely suggested as a possible alternative to cow milk in allergic subjects, because of the high genetic variability in alpha-s1 casein (CSN1S1) content, which is associated with different technological and nutritional properties of milk. The aim of the study was to evaluate the suitability of goat milk with low and high CSN1S1 to produce fermented milk. In addition, the performance as starter of selected Lactobacillus paracasei FS109 strain compared to no-selected L. paracasei strains was investigated. Initially, the selected L. paracasei FS109 strain was tested for adhesion ability to HT-29 and Caco-2 cells and immunomodulation effect. Then, the strain was used to produce fermented milk from goat milk with a low and high casein CSN1S1 genotype. The results indicated that greater acidifying activity was obtained for L. paracasei FS109 after 24 h of fermentation than the other two strains tested independently by the CSN1S1 genotype. L. paracasei FS109 grew well during fermentation, reaching a higher value (>8.5 log CFU/mL). Interestingly, the same strain maintained a high viable population (about 9 log CFU/mL) during the 30-day cold storage of the product. The present study shows for the first time the suitability of the goat milk with low CSN1S1 genotypes to produce fermented milk and highlight the importance of strain selection in determination of technological and beneficial traits. Combining goat milk with low CSN1S1 and selected strains could be a strategy of improving traditional and functional fermented milk market. Full article

In the past, some microbiological studies have considered most non-Saccharomyces species to be undesirable spoilage microorganisms. For several decades, that belief made the Saccharomyces genus the only option considered by winemakers for achieving the best possible wine quality. Nevertheless, in recent decades, some strains of non-Saccharomyces species have been proven to improve the quality of wine. Non-Saccharomyces species can positively influence quality parameters such as aroma, acidity, color, and food safety. These quality improvements allow winemakers to produce innovative and differentiated wines. For that reason, the yeast strains Torulaspora delbrueckii, Lachancea thermotolerans, Metschnikowia pulcherrima, Schizosaccharomyces pombe, and Pichia kluyveri are now available on the market. Other interesting species, such as Starmerella bacillaris, Meyerozyma guilliermondii, Hanseniospora spp., and others, will probably be available in the near future. Full article